Sirs and Madams,
In January I posted a query about digitizing the counting of
worms on 60mm
agar plates, and I received some helpful replies regarding potential image
analysis software (thanks to all who replied). However, it has become
apparent that the difficulty is not with image analysis software per se,
but rather with the quality of the image we can get with our
camera/microscope setup. We are currently using a Diagnostic Instruments
RT Color Spot Model 2.2.0 digital camera with the Spot Image grabbing
software, and we cannot get sufficiently good images at low magnification
(20X) to reliably count all the worms on the plate.
The problem to be solved is this: we need to ACCURATELY count dead (or
immobilized) worms of all stages (>= L1) on a 60 mm agar plate, and we need
very high throughput. A trained observer can currently count by eye all
the worms on a plate in approximately 3 minutes. The advantages of
automating this process are obvious. If anyone has devised (or can think
of) a solution to this particular problem, we will be most grateful if you
share it with us.
One possibility that has been suggested is a stain that will
stain worms
but not E. coli. I haven't come across such a stain, but again,
suggestions will be most appreciated.
Thanks,
-Charlie Baer
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Charles F. Baer
Ecology and Evolution Program
1210 University of Oregon
Eugene, OR 97403-1210 USA
Phone: 541-346-4697
FAX: 541-346-2364
Email: cbaer at darkwing.uoregon.edu
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